CN114010695A - Traditional Chinese medicine formula for resisting animal dermatomycosis and preparation method - Google Patents
Traditional Chinese medicine formula for resisting animal dermatomycosis and preparation method Download PDFInfo
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
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- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/75—Rutaceae (Rue family)
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- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/75—Rutaceae (Rue family)
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- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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Abstract
The invention relates to the technical field of animal dermatomycosis treatment medicines, and discloses a traditional Chinese medicine formula for resisting animal dermatomycosis, which comprises the following components: radix Sophorae Flavescentis, cortex Dictamni Radicis, cortex Phellodendri, cortex pseudolaricis and fructus Cnidii; also discloses a preparation method. The formula of the invention has obvious effect on animal source trichosporium, microsporum gypseum, candida albicans and the like, can achieve the treatment aim, basically has no toxic or side effect, and can reduce the drug resistance.
Description
Technical Field
The invention relates to the technical field of animal dermatomycosis treatment medicines, in particular to a traditional Chinese medicine formula for resisting animal dermatomycosis and a preparation method thereof.
Background
The fungal dermatosis is a zoonosis caused by dermatophyte, and is commonly called as copper ringworm, tinea capitis, tinea moubata, psoriasis and the like. After the animals are infected with fungi, the skin is itchy, anorexia, emaciation and immunity are reduced, so that the nutrient consumption is increased, the labor is wasted, the breeding cost is increased, the profit is suddenly reduced, and great economic loss is caused to a breeding (household) farm. The wild animals are more difficult to radically cure after being infected with dermatophytosis, which particularly affects the appearance and the public image and causes serious influence on the tourism industry. Wherein the Trichosporon is one of susceptible fungi of beef cattle and wild animals. The currently used antifungal drugs are easy to cause administration difficulty, have toxicity, and easily cause side effects such as drug resistance of fungi and the like.
Disclosure of Invention
Based on the problems, the invention provides a traditional Chinese medicine formula for resisting animal dermatomycosis and a preparation method thereof, and the traditional Chinese medicine formula has obvious effect on animal dermatomycosis such as animal trichosporium, microsporidia gypseum, candida albicans and the like, and can achieve the aim of treatment.
In order to solve the technical problems, the invention provides a traditional Chinese medicine formula for resisting animal dermatomycosis, which comprises the following components: radix Sophorae Flavescentis, cortex Dictamni Radicis, cortex Phellodendri, cortex pseudolaricis and fructus Cnidii.
Further, the weight parts of each component are as follows: 5-20 parts of radix sophorae flavescentis, 10-30 parts of cortex dictamni, 30-50 parts of cortex phellodendri, 5-20 parts of pseudolarix and 20-40 parts of fructus cnidii.
Further, the weight parts of each component are as follows: 7.92 parts of radix sophorae flavescentis, 17.5 parts of cortex dictamni, 39.7 parts of golden cypress, 12 parts of pseudolarix and 22.9 parts of fructus cnidii.
Further, the weight parts of each component are as follows: 23.4 parts of lightyellow sophora root, 6.05 parts of cortex dictamni, 48.9 parts of golden cypress, 3.37 parts of pseudolarix and 18.2 parts of fructus cnidii.
Further, the weight parts of each component are as follows: 14.6 parts of lightyellow sophora root, 11.2 parts of cortex dictamni, 44.9 parts of amur corktree bark, 22.9 parts of pseudolarix and 6.42 parts of common cnidium fruit.
In order to solve the technical problems, the invention also provides application of the traditional Chinese medicine formula in an animal antifungal medicine.
Further, the fungi include animal-derived trichosporium, microsporidia gypseum, and candida albicans.
In order to solve the technical problems, the invention also provides a preparation method of the traditional Chinese medicine formula, which comprises the following steps:
s1: pulverizing the medicinal materials in the Chinese medicinal formula respectively with a Chinese medicinal pulverizer, sieving with a 60 mesh sieve, placing in a white tray, drying at 60 deg.C overnight for use, and drying to constant weight;
s2: accurately weighing the medicinal powder of different medicinal material components in the step S1 in proportion, uniformly mixing, soaking in ethanol for 3 hours, wherein the material-liquid ratio of the medicinal powder to the ethanol is 1: 12, carrying out ultrasonic reaction for 40min under the ultrasonic condition at the reaction temperature of 60 ℃ under the ethanol concentration of 70 percent to finish extraction to obtain an extracting solution;
s3: collecting the extractive solution obtained in step S2, concentrating at 60 deg.C under reduced pressure to 8 kilopascals by using rotary evaporator until each milliliter of medicinal liquid contains 1g of crude drug to obtain concentrated solution;
s4: performing vortex oscillation on the concentrated solution in the step S3 at normal temperature for 5min, adding 2% dimethyl sulfoxide, performing constant volume to 10mL, performing oscillation and shaking again, and then putting into a sterile centrifuge tube for freezing preservation at-20 ℃ for later use;
s5: when the concentrated solution is clinically used, the concentrated solution obtained in the step S4 is diluted by 10-100 times;
s6: or making into unguent, oil and spray according to the situation.
Further, the ultrasonic frequency in step S2 is 40 KHz.
Compared with the prior art, the invention has the beneficial effects that: the formula of the invention has obvious effect on animal source trichosporium, candida albicans, microsporopsis gypseum and the like, can achieve the treatment aim, basically has no toxic or side effect, and can reduce the drug resistance.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not used as limitations of the present invention.
Example (b):
the test apparatus used in this example was as follows: microplate reader MK3 tib: THERMO FISH; centrifuge instetat TG 16: changsha Yingtai instruments, Inc.; incubator DH 3600A: tianjin city apparatus of the delight; an ultrasonic oscillator; a rotary evaporator; high-pressure steam sterilizing pot. The test reagents used were as follows: xtt; menadione sodium bisulfite; PBS buffer solution; dimethyl sulfoxide; acetone; and (3) ethanol.
The embodiment designs a traditional Chinese medicine formula for resisting animal dermatomycosis, which comprises the following components: radix Sophorae Flavescentis, cortex Dictamni Radicis, cortex Phellodendri, cortex pseudolaricis and fructus Cnidii.
As shown in table 1, in this embodiment, a uniform design model of a traditional Chinese medicine formula is adopted, an experiment is designed by a uniform design method of the formula, and 16 levels are selected from 3.0 uniform design of software to establish 5-factor 16-level 16-experiment unlimited uniform design of the formula.
TABLE 1U16(165) Uniform design table
In this embodiment, the medicinal materials in the formula of the above traditional Chinese medicines are respectively pulverized by a traditional Chinese medicine pulverizer, and then screened by a 60-mesh screen, and then contained in a white tray, and dried overnight at 60 ℃ for later use, and the medicine is dried to constant weight. And then accurately weighing the medicinal powder of different medicinal material components in proportion, uniformly mixing, and soaking in ethanol for 3 hours, wherein the material-liquid ratio of the medicinal powder to the ethanol is 1: 12, carrying out ultrasonic reaction for 40min under the ultrasonic condition with the ethanol concentration of 70 percent and the ultrasonic frequency of 40KHz and the reaction temperature of 60 ℃ to finish extraction, collecting an extracting solution, reducing the pressure to 8 kilopascals by using a rotary evaporator under the condition of 60 ℃ to carry out concentration until 1g of the crude drug is contained in each milliliter of the liquid medicine. Concentrating, collecting test liquid, subjecting the test liquid to vortex shaking at room temperature for 5min, adding 2% dimethyl sulfoxide, diluting to constant volume of 10mL, centrifuging, collecting supernatant, placing into a sterile centrifuge tube, and freezing at-20 deg.C for storage, wherein the prepared solution is diluted by 10-100 times.
The biological quilt inhibition rate of the traditional Chinese medicine formula on the animal-derived trichosporium is researched by adopting an XTT reduction method according to the following steps. Inoculating bovine-derived spore strain preserved at-20 deg.C to SDA medium, growing at 32 deg.C and 5% CO2Culturing for 48h, placing the strain in SDB culture medium, shaking and culturing at 30 deg.C and 180r/min for 12h, filtering with microporous membrane to collect cells of bovine-derived Trichosporon in logarithmic growth state, adding SDB culture medium, mixing, and preparing into the final product with concentration of 1 × 10 by hemocytometer6one/mL of standard bacterial suspension. And (3) inoculating 100 mu L of standard bacterial suspension into a 96-hole microplate, adding 100 mu L of sdb culture medium, incubating at 32 ℃ for 36h, and washing 3 times by 100 mu L of PBS per hole to finish the preparation of the bovine-derived trichosporium biofilm.
After the biofilm is prepared, the activity of the traditional Chinese medicine formula for inhibiting the bovine-derived trichosporium tunica is determined, the bovine-derived trichosporium is continuously cultured for 24 hours by using SDB culture media containing traditional Chinese medicine formula preparations (mu mol/L) with different concentrations respectively, and a negative control group without the traditional Chinese medicine formula preparation and a blank control group without the bovine-derived trichosporium are established. Using PBS solution as solution, preparing XTT into 0.5mg/mL solution, filtering with 0.22 μm microporous membrane, subpackaging, and storing at-20 deg.C in dark. Using menadione to prepare 12mmol/L solution, subpackaging, and storing at-20 ℃ in the dark. Preparing an XTT/menadione solution before use to ensure that the final concentration of menadione is 1.2 mu mmol/L, respectively adding 100 mu L of the XTT/menadione solution into each hole, carrying out a light-shielding reaction at 37 ℃ for 3h, and reading the absorbance value (A) at the wavelength of 490nm of each hole by using a microplate reader after the reaction is finished.
The relative inhibition (Y) per well was calculated according to the formula Y ═ 1- { experimental group-blank control group/negative control group-blank control group } × 100%, the greater the Y value, the greater the inhibition intensity. The lowest concentration (SMIC50) of the traditional Chinese medicine compound for inhibiting the activity of 50 percent of the bovine-derived trichotheca is the lowest effective concentration of the traditional Chinese medicine compound when Y is more than 50 percent. The experiment was repeated 3 times, 3 replicate wells were set for each concentration, and the average was taken. Experimental data are presented as mean ± standard deviation using t-significance test and multiple comparisons between groups. The results are shown in table 2, and it can be seen that the inhibition rate of the formulation No. 12 at the corresponding concentration is up to 99.62%, the effect is extremely excellent, the effect is comparable to that of the positive drug fluconazole, and the cost performance is high.
TABLE 2 biological quilt inhibition of bovine-derived Trichosporon at different concentrations for different Chinese medicinal compositions
This example also examined the therapeutic effect of the above formulation on candida albicans, and the MIC test effect is shown in the following table, wherein-: the inhibition effect is obvious, and the solution is clear; +: the inhibition effect is not obvious, and the solution is turbid.
MIC test Effect (Effect on Candida albicans)
The results in the table above show that: all the raw liquid of the formula can inhibit the bacteria; the formulas 9 and 12 have the best effect, and can completely inhibit the growth of the bacteria after being diluted by 1000 times; the effects of the formulas 3 and 6 are good, and the effect of 100 times dilution is good.
Through experiments, the finally determined traditional Chinese medicine formula for resisting the animal dermatophytes comprises the following components in parts by weight: 5-20 parts of radix sophorae flavescentis, 10-30 parts of cortex dictamni, 30-50 parts of cortex phellodendri, 5-20 parts of pseudolarix and 20-40 parts of fructus cnidii. The traditional Chinese medicine formula can be applied to animal antifungal dermatological drugs, and the fungi comprise animal source trichosporium, microsporum gypseum and candida albicans. The traditional Chinese medicine formula can be prepared into various dosage forms such as paste, spray and the like, and the dosage forms can be selected according to requirements in practical application.
In the embodiment, the traditional Chinese medicine formula is applied to the treatment of cattle with confirmed fungal skin diseases in actual clinic, and currently 63 cattle with confirmed fungal skin diseases are mainly treated as follows: 47 cattle in rural agricultural bureau of Yuexi county, continuously spraying for 2 times a day, and eliminating clinical symptoms after continuously using for one week; continuously spraying 10 cattle, ten thousand cattle husbandry and Limited liability company in Vietnam county, 2 times a day, and eliminating clinical symptoms after one week; 6 cattle of farmer professional cooperative are cultivated in Yuetui county in Yuetui province, and are treated by continuous spraying 2 times a day, and clinical symptoms disappear after one week. The traditional Chinese medicine formula of the embodiment is applied to multiple beef cattle farms in Yanyuan county to treat more than 100 beef cattle, the clinical application effect is good, and clinical symptoms disappear after continuous spray treatment for one week and 2 times a day. The formula of the embodiment is also applied to dog and cat fungal skin disease treatment by multiple pet hospitals, and the clinical symptoms disappear after one week after continuous spray treatment and 2 times a day. China panda protection panda research center 7 pandas suffering from fungal dermatosis use the continuous spray treatment formula of the embodiment for 2 times a day, and clinical symptoms disappear after one week. The formula of the embodiment can also be prepared into ointment, and the using effect is good.
The above is an embodiment of the present invention. The embodiments and specific parameters in the embodiments are only for the purpose of clearly illustrating the process of verifying the invention and are not intended to limit the scope of the invention, which is defined by the claims, and all the equivalent structural changes made by applying the content of the specification of the invention should be covered by the scope of the invention.
Claims (10)
1. The traditional Chinese medicine formula for resisting animal dermatomycosis is characterized by comprising the following components: radix Sophorae Flavescentis, cortex Dictamni Radicis, cortex Phellodendri, cortex pseudolaricis and fructus Cnidii.
2. The traditional Chinese medicine formula for resisting animal dermatomycosis according to claim 1, which is characterized by comprising the following components in parts by weight: 5-20 parts of radix sophorae flavescentis, 10-30 parts of cortex dictamni, 30-50 parts of cortex phellodendri, 5-20 parts of pseudolarix and 20-40 parts of fructus cnidii.
3. The traditional Chinese medicine formula for resisting animal dermatomycosis according to claim 2, is characterized in that the weight parts of the components are as follows: 7.92 parts of radix sophorae flavescentis, 17.5 parts of cortex dictamni, 39.7 parts of golden cypress, 12 parts of pseudolarix and 22.9 parts of fructus cnidii.
4. The traditional Chinese medicine formula for resisting animal dermatomycosis according to claim 2, is characterized in that the weight parts of the components are as follows: 23.4 parts of lightyellow sophora root, 6.05 parts of cortex dictamni, 48.9 parts of golden cypress, 3.37 parts of pseudolarix and 18.2 parts of fructus cnidii.
5. The traditional Chinese medicine formula for resisting animal dermatomycosis according to claim 2, is characterized in that the weight parts of the components are as follows: 14.6 parts of lightyellow sophora root, 11.2 parts of cortex dictamni, 44.9 parts of amur corktree bark, 22.9 parts of pseudolarix and 6.42 parts of common cnidium fruit.
6. The use of a herbal formulation according to any one of claims 1 to 5 in the preparation of an antifungal medicament for animals.
7. The use according to claim 6, wherein the fungi comprise animal-derived Trichosporon, Microsporum gypseum and Candida albicans.
8. The method for preparing a traditional Chinese medicine formula according to any one of claims 1 to 5, which is characterized by comprising the following steps:
s1: pulverizing the medicinal materials in the Chinese medicinal formula respectively with a Chinese medicinal pulverizer, sieving with a 60 mesh sieve, placing in a white tray, drying at 60 deg.C overnight for use, and drying to constant weight;
s2: accurately weighing the medicinal powder of different medicinal material components in the step S1 in proportion, uniformly mixing, soaking in ethanol for 3 hours, wherein the material-liquid ratio of the medicinal powder to the ethanol is 1: 12, carrying out ultrasonic reaction for 40min under the ultrasonic condition at the reaction temperature of 60 ℃ under the ethanol concentration of 70 percent to finish extraction to obtain an extracting solution;
s3: collecting the extractive solution obtained in step S2, concentrating at 60 deg.C under reduced pressure to 8 kilopascals by using rotary evaporator until each milliliter of medicinal liquid contains 1g of crude drug to obtain concentrated solution;
s4: and (4) carrying out vortex oscillation on the concentrated solution in the step S3 for 5min at normal temperature, adding 2% dimethyl sulfoxide, fixing the volume to 10mL, oscillating and shaking uniformly again, and then putting into a sterile centrifuge tube for freezing preservation at-20 ℃ for later use.
9. The method according to claim 8, wherein the ultrasonic frequency in step S2 is 40 KHz.
10. The method according to claim 9, wherein the solution prepared after the treatment of step S4 is diluted 10-100 times at the time of use, and the solution prepared after the treatment of step S4 is formulated into paste, oil and spray.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202111359256.8A CN114010695A (en) | 2021-11-17 | 2021-11-17 | Traditional Chinese medicine formula for resisting animal dermatomycosis and preparation method |
| ZA2022/08569A ZA202208569B (en) | 2021-11-17 | 2022-08-01 | Traditional chinese medicine formula for resisting animals dermatomycosis and preparation method thereof |
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| CN202111359256.8A CN114010695A (en) | 2021-11-17 | 2021-11-17 | Traditional Chinese medicine formula for resisting animal dermatomycosis and preparation method |
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2021
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2022
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